Quantitative analysis of leukocyte phagcytosis in 100 µl of whole blood under optimal conditions without any disturbance by isolation procedures A. Sample Preparation for opsonized E.coli-FITC. Simple as you know it from immunofluorescence analysis: 1.Labeling 2. Quenching 3. Lysing and Fixation 4. Washing and DNA Staining B. Measurement Gating on leukocytes after DNA staining: Discrimination of bacteria, single leukocytes and doublets C. Data Evaluation Gate on monocytes and granulocytes and determine the percentage of phagocytosing cells and their phagocytic activity (= mean fluorescence = number of bacteria per cell) Report your results

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